Codon modified amylase from Bacillus akibai

Abstract

The present invention relates to an isolated synthetic polynucleotide encoding the mature amylase AX856 from Bacillus akibai , using codon modified polynucleotide constructs for the expression of the amylase.

The invention claimed is: 
     
       1. An isolated synthetic polynucleotide encoding a mature amylase, said polynucleotide comprising the nucleotide sequence shown in SEQ ID NO:4. 
     
     
       2. The isolated synthetic polynucleotide of  claim 1  which consists of the nucleotide sequence shown in SEQ ID NO:4. 
     
     
       3. A polynucleotide construct comprising a polynucleotide of  claim 2  operably linked to control sequences that provide for its expression in a host cell of choice. 
     
     
       4. The polynucleotide construct of  claim 3 , wherein the control sequences comprise a polynucleotide region that encodes a signal peptide fused to the N-terminus of the amylase which directs the amylase into the secretory pathway of the host cell. 
     
     
       5. The polynucleotide construct of  claim 4 , wherein the signal peptide is the amyL signal peptide. 
     
     
       6. A polynucleotide construct comprising a polynucleotide of  claim 1  operably linked to control sequences that provide for its expression in a host cell of choice. 
     
     
       7. The polynucleotide construct of  claim 6 , wherein the control sequences comprise a tandem promoter. 
     
     
       8. The polynucleotide construct of  claim 7 , wherein the control sequences comprise a polynucleotide region that encodes a signal peptide fused to the N-terminus of the amylase which directs the amylase into the secretory pathway of the host cell. 
     
     
       9. The polynucleotide construct of  claim 8 , wherein the signal peptide is the amyL signal peptide. 
     
     
       10. A recombinant host cell transformed with or comprising a polynucleotide of  claim 1 , wherein said host cell produces the mature amylase. 
     
     
       11. The host cell of  claim 10 , which is a  Bacillus  cell. 
     
     
       12. The host cell of  claim 11 , which is a  Bacillus licheniformis  cell. 
     
     
       13. The host cell of  claim 11 , which is a  Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillus brevis, Bacillus circulans, Bacillus clausii, Bacillus coagulans, Bacillus firmus, Bacillus lautus, Bacillus lentus, Bacillus megaterium, Bacillus pumilus, Bacillus stearothermophilus, Bacillus subtilis , or  Bacillus thuringiensis  cell. 
     
     
       14. A method for producing a mature amylase, comprising cultivating a host cell of  claim 10  under conditions conducive for the expression of the amylase. 
     
     
       15. The method of  claim 14 , further comprising recovering the amylase. 
     
     
       16. A method for producing a mature amylase, comprising cultivating a host cell of  claim 11  under conditions conducive for the expression of the amylase. 
     
     
       17. The method of  claim 16 , further comprising recovering the amylase. 
     
     
       18. A method for producing a mature amylase, comprising cultivating a host cell of  claim 12  under conditions conducive for the expression of the amylase. 
     
     
       19. The method of  claim 18 , further comprising recovering the amylase. 
     
     
       20. A method for producing a mature amylase, comprising cultivating a host cell of  claim 13  under conditions conducive for the expression of the amylase.